Cooperativity of concanavalin A patching and its influence on cytoskeleton changes in fibroblast rounding and detachment.
We have studied the extent of concanavalin A (Con A) patching on spread fibroblasts as a function of increasing concentration of added Con A, and also the influence of this patching on subsequent cytoskeleton changes during exposure to trypsin and EGTA. Living cells do not patch at low concentrations of Con A (around 1 microgram ml-1) but progressively do so at higher levels (2-10 microgram ml-1), in very close parallel with a cooperative phase of binding characterized earlier. The proportion of visibly patched cells is at a maximum when this cooperative phase is complete and the correlation remains when cells are treated with azide of cytochalasin B to shift the concentration range over which both phenomena occur. All this evidence points to a common origin for cooperative binding and visible patching which is sensitive to the state of cellular actomyosin, and involves the mediation of actin-associated structures. As a consequence of Con A patching, cell attachment is preserved against the action of EGTA. On patched cells, this agent causes minor if any changes in cell shape and in the distributions of microtubules and 10-nm filaments, but actin stress fibres are dispersed. However, conditions appear to be retained for reassembly of the stress fibres because these can be caused to reappear within minutes after removal of EGTA when glucose, or Ca2+ or Mn2+ in the absence of glucose, is supplied in the medium. Trypsin caused only partial destruction of stress fibres in Con A-treated cells and did not promote detachment or dispersal of microtubules or 10-nm filament systems.
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002642:Chick Embryo; D003208:Concanavalin A / Q000378:metabolism*; D006224:Cricetinae; D004533:Egtazic Acid / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D007668:Kidney / Q000166:cytology; D051381:Rats; D014357:Trypsin / Q000494:pharmacology
D003208:Concanavalin A; D004533:Egtazic Acid; D014357:Trypsin